Calditerrivibrio Medium (DSMZ Medium 1112)

[所属分类:培养基配方] [发布时间:2020-3-26] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Calditerrivibrio Medium
(DSMZ Medium 1112)

培养基配方
Composition per liter:
NaNO3  ........................................................0.85g
Na-acetate ....................................................0.82g
NH4Cl .........................................................0.54g
MgCl2 ·6H20....................................................0.2g
CaCl2 ·2H2O...................................................0.15g
KH2PO4 ........................................................0.14g
Resazurin .....................................................0.5mg
NaHCO3 solution...............................................10.0mL
Vitamin solution..............................................10.0mL
Na2S·9H2O solution...........................................10.0mL
Trace element solution SL-10...................................1.0mL
Selenite/tungstate solution ...................................1.0mL
pH 7.0 ± 0.2 at 25°C
NaHCO3 Solution :
Composition per 10.0mL:
NaHCO3 ........................................................2.5g
Preparation of NaHCO3 Solution:  Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Sparge
with 20% CO2 + 80% H2 . Autoclave for 15 min at 15 psi pressure–
121°C. Cool to room temperature.
Na2S·9H2O Solution :
Composition per 10.0mL:
Na2S·9H2O.....................................................0.5g
Preparation of Na2S·9H2O Solution: Add Na2S·9H2O to dis-
tilled/deionized water and bring volume to 10.0mL. Mix thoroughly.
Autoclave under 100% N2 for 15 min at 15 psi pressure–121°C. Cool
to room temperature.
Selenite/Tungstate Solution:
Composition per liter:
NaOH.........................................................0.5g
Na2WO4 ·2H2O...............................................4.0mg
Na2SeO3 ·5H2O..............................................3.0mg
Preparation of Selenite/Tungstate Solution:  Add components
to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–121°C.
Vitamin Solution:
Composition per liter:
Pyridoxine-HCl..............................................10.0mg
Thiamine-HCl·2H2O...........................................5.0mg
Riboflavin ..................................................5.0mg
Nicotinic acid...............................................5.0mg
D -Ca-pantothenate...........................................5.0mg
p-Aminobenzoic acid..........................................5.0mg
Lipoic acid .................................................5.0mg
Biotin ......................................................2.0mg
Folic acid...................................................2.0mg
Vitamin B 12 ................................................0.1mg
Preparation of Vitamin Solution:  Add components to distilled/
deionized water and bring volume to 1.0L. Mix thoroughly. Sparge
with 80% H2 + 20% CO2 . Filter sterilize.
Trace Elements Solution SL-10:
Composition per liter:
FeCl2 ·4H2O..................................................1.5g
CoCl2 ·6H2O...............................................190.0mg
MnCl2 ·4H2O ..............................................100.0mg
ZnCl2 ......................................................70.0mg
Na2MoO4 ·2H2O..............................................36.0mg
NiCl2 ·6H2O................................................24.0mg
H3BO3 .......................................................6.0mg
CuCl2 ·2H2O.................................................2.0mg
HCl (25% solution)..........................................10.0mL
Preparation of Trace Elements Solution SL-10: Add FeCl2 ·4H2O
to 10.0mL of HCl solution. Mix thoroughly. Add distilled/deionized
water and bring volume to 1.0L. Add remaining components. Mix thor-
oughly. Sparge with 100% N2 . Autoclave for 15 min at 15 psi pressure–
121°C.
Preparation of Medium: Add components, except bicarbonate so-
lution, sulfide solution, and vitamin solution, to distilled/deionized wa-
ter and bring volume to 970.0mL. Mix thoroughly. Gently heat and
bring to boiling. Boils for several minutes. Cool to room temperature
while sparging with 80% N2 + 20% CO2 . Distribute into screw-capped
tubes or bottles under an atmosphere of 80% N2 + 20% CO2 . Autoclave
for 15 min at 15 psi pressure–121°C. Add the bicarbonate solution, sul-
fide solution, and vitamin solution. Adjust the final pH to 7.0.
Use:  For the cultivation of Calditerrivibrio spp.